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The results of PVA can then be exported to spreadsheet software a

The results of PVA can then be exported to spreadsheet software and statistically analysed. The PVA function has now been fully integrated into the GRaPe software. 3. Results 3.1. The Genome-Scale Kinetic Model of Mycobacterium Tuberculosis There are now several genome-scale metabolic reconstructions of M. tuberculosis [15,19,26] which can serve as a basis to construct a genome-scale kinetic model. In [15] the authors constructed a genome-scale metabolic network of M. tuberculosis using a reconstruction of Streptomyces coelicolor as a starting point. Genes were mapped between the two Inhibitors,research,lifescience,medical species using gene orthology clusters from the Kyoto Encyclopaedia of Genes and Genomes (KEGG) [27]. Using the

KEGG and BioCyc databases Inhibitors,research,lifescience,medical and

analysis of relevant research articles, the authors further supplemented the initial model. The final metabolic network of M. tuberculosis includes this website reactions needed for the synthesis of the cell membrane, complex lipids and carbohydrates, which are important for both growth and pathogenesis. Other metabolic pathways Inhibitors,research,lifescience,medical such as respiratory pathways and synthesis of biomolecules, which are specific to mycobacteria, were also modelled manually, as well as iron metabolism and transport reactions responsible for the import of carbon, nitrogen, minerals and compounds of high molecular weight. The final stoichiometric model reconstructed by the Beste group consists of 739 metabolites, 849 reactions and 726 genes; Inhibitors,research,lifescience,medical they calibrated their model by growing Mycobacterium bovis bacilli Calmette Guérin in a continuous culture and measuring parameters for steady-state growth. FBA was used to calculate substrate consumption rates. Their results showed a close agreement with experimentally determined Inhibitors,research,lifescience,medical values. The model was made available as a web-based interactive tool. Using GRaPe [7], we created a genome-scale kinetic model of M. tuberculosis based on the stoichiometric model developed by Beste et al.

[15]. GRaPe assigned an enzyme species to each reaction, which was then mapped to the corresponding gene(s) provided Carnitine dehydrogenase in the original reconstruction. All reactions were assumed to follow a random-order mechanism. We then used GRaPe to generate generic rate equations for all the reactions in the M. tuberculosis genome-scale network. The type of rate equation generated for each reaction was based on the stoichiometry of the reaction, as described in the Methods section. The resulting genome-scale model of M. tuberculosis contains 739 metabolites, 856 metabolic reactions and 856 enzyme species. 3.2. Parameter Estimation We obtained flux distributions for three steady-states with glycerol being the only carbon source using the interactive web-based tool developed by Beste et al. [15]. The tool uses FBA to calculate flux distributions for the three steady-states with glycerol consumption at 0, 0.